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BACKGROUND: Wound healing monitoring and timely decision-making are critical for wound classification. Tryptophan (Tr) intrinsic fluorescence, detected at 295/340 nm, provides a noninvasive approach for wound assessment. Our previous work demonstrated that this autofluorescence is associated with keratinocytes in a highly proliferative state in vitro. OBJECTIVE: We investigated the correlation between Tr fluorescence and key wound healing parameters, including re-epithelialization, fibrosis, neovascularization, and acute and chronic inflammation, using a rabbit model. METHODS: Seven rabbits underwent wound healing assessment over a 15-day period. We employed histological analysis from central and marginal biopsies, and UV fluorescence imaging captured by a monochromatic near-UV sensitive camera equipped with a passband optical filter (340 nm/12 nm). Excitation was achieved using a 295 nm LEDs ring lamp. Normalized fluorescence values were correlated with histological measurements using Pearson correlation. RESULTS: The UV fluorescence strongly exhibited a strong correlation with re-epithelization (r = 0.8) at the wound edge, with peak intensity observed between the sixth and ninth days. Notably, wound-healing dynamics differed between the wound center and edge, primarily attributed to variations in re-epithelialization, neovascularization, and chronic inflammation. CONCLUSION: Our findings highlight the presence of autofluorescence at 295/340 nm during wound healing, demonstrating a robust association with re-epithelialization. This excitation/emission signal holds promise as a valuable noninvasive strategy for monitoring wound closure, re-epithelialization, and other biological processes where Tr plays a pivotal role.
Assuntos
Reepitelização , Triptofano , Cicatrização , Animais , Coelhos , Reepitelização/fisiologia , Cicatrização/fisiologia , Modelos Animais de Doenças , Fluorescência , Pele/patologia , Pele/lesões , Imagem Óptica/métodos , Inflamação/patologia , Raios UltravioletaRESUMO
Build-up of extracellular matrix in liver fibrosis results in changes on endogenous molecules expression that may be studied through the fluorescence characterization of ex vivo liver samples. To the best of our knowledge, no investigations have provided in-depth evidence and discussion on the changes of the endogenous fluorescence in ex vivo tissue due to the effects of the preservation media. In this work, we contrast and analyze the endogenous fluorescence from tryptophan, vitamin A, hydroxyproline and elastin cross-links potential biomarkers of the liver fibrosis, in in vivo measurements and liver samples preserved on formaldehyde, and two standard preservation media. As it is known, chemical changes in tissue, caused by formaldehyde fixation, alter the endogenous fluorescence spectra. We propose the use of phosphate-buffered saline (PBS), and Iscove's Modified Dulbecco's Medium (IMDM) to elude the fluorescence changes. PBS and IMDM showed to maintain the endogenous fluorescence characteristics similar to in vivo conditions. The results of this work point the way for a more reliable assessment of endogenous fluorescence in ex vivo hepatic studies.
Assuntos
Fluorescência , Hidroxiprolina/análise , Cirrose Hepática/diagnóstico , Triptofano/análise , Vitamina A/análise , Biomarcadores/análise , Meios de Cultura/química , Formaldeído , Humanos , Cirrose Hepática/induzido quimicamente , Fosfatos/química , Cloreto de Sódio/química , Espectrometria de FluorescênciaRESUMO
BACKGROUND: High alcohol intake on weekends (binge drinking) is more frequent in young adults, who could undergo early liver damage. Alcohol-induced liver damage is characterized by polymorphonuclear cell (PMN) infiltration, which can be represented in the peripheral blood by altered trafficking and activation profiles. OBJECTIVE: To evaluate the PMN trafficking and activation immunophenotypic profiles in people with a binge drinking pattern. METHODS: People with binge drinking (n = 18, 8 females) or at low risk (n = 16, 13 females) based on their AUDIT and HEPCA scores were studied. Hematic biometry and liver enzyme tests were conducted. Peripheral blood leukocytes were stained for CCR5, CCR4, and CXCR4 (trafficking) and CD69 and CD127 (activation). PMNs and monocytes were analyzed by FACS. The data were analyzed using the T-test and Mann-Whitney's U-test for contrasts and principal component and Fuzzy C means analyses for clustering, with p < 0.05 considered significant. RESULTS: Compared to the low-risk group, the binge group showed higher CCR5 expression on PMNs, decreases in the CD69 percentage and positive PMNs per microliter, and decreased CXCR4 expression on monocytes. Six immunophenotypical clusters were identified, all of which were distributed following the CCR5 and CXCR4 main vectors. CONCLUSION: Young adult binge drinkers have differential PMN trafficking and activation immunophenotypes, which could be related to the initial onset of alcoholic liver disease and a systemic inflammatory state in response to their alcohol consumption pattern. These findings could lead to the future development of an early diagnostic tool.
Assuntos
Consumo Excessivo de Bebidas Alcoólicas/sangue , Leucócitos Mononucleares/imunologia , Monócitos/imunologia , Adulto , Consumo Excessivo de Bebidas Alcoólicas/imunologia , Feminino , Humanos , Imunofenotipagem , Masculino , Adulto JovemRESUMO
Background: Robot-assisted radical prostatectomy has positioned itself as the approach of choice in the treatment of prostate cancer. Objective: To compare the outcomes of robot-assisted radical prostatectomy using the Retzius-Sparing (RS) approach against the modified Frankfurt (MF) technique. Method: To describe the perioperative, functional and oncological outcomes of 13 patients with prostate cancer who underwent RS robotic radical prostatectomy compared to MF, evaluating pathological results, urinary continence, sexual function and oncological control in 1 year of follow-up. Results: The average age was 64 years in RS group vs. 61 years in MF group. The values of total prostate antigen were higher in the RS group (25 ng/dl) vs. MF group (11 ng/dl). The volume of gland in RS group was 40.62 ml vs. 63.33 ml in the RS group. All patients were bilaterally neuropreserved, being statistically significant in favor of MF group (p = 0.016). Positive surgical margins were higher in R-S group (38.4%) vs. MF group (33.3%). Conclusions: With RS the same tendency to urinary continence is observed, with a significant difference in erectile function in favor of MF. This preliminary study shows better impact on erectile function.
Antecedentes: La prostatectomía radical asistida por robot se ha posicionado como el abordaje de elección en el tratamiento del cáncer de próstata. Objetivo: Comparar los resultados de la prostatectomía radical asistida por robot utilizando el abordaje Retzius-Sparing (RS) contra el Frankfurt modificado (FM). Método: Se describen los desenlaces perioperatorios, funcionales y oncológicos de 13 pacientes con cáncer de próstata que fueron llevados a prostatectomía radical robótica con RS, en comparación con FM, y se evalúan los resultados patológicos, continencia urinaria, función sexual y control oncológico a 1 año de seguimiento. Resultados: La media de edad fue de 64 años en el grupo RS y de 61 años en el grupo FM. Los valores de antígeno prostático total fueron mayores en el grupo RS (25 ng/dl) que en el FM (11 ng/dl). El volumen de la glándula fue menor en el grupo RS (40.62 ml) que en el FM (63.33 ml). Todos los pacientes fueron neuropreservados bilateralmente, siendo la diferencia estadísticamente significativa a favor de FM (p = 0.016). Los márgenes quirúrgicos positivos fueron mayores en el grupo RS (38.4%) que en el FM (33.3%). Conclusiones: Con RS se observa la misma tendencia a la continencia urinaria, con diferencia significativa en la función eréctil a favor de FM. Este estudio preliminar muestra mejor impacto en la función eréctil.
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Introduction: The enzyme lactate dehydrogenase (LDH) is a good marker of general hyperinflammation correlated with mortality for COVID-19, and is therefore used in prognosis tools. In a current COVID-19 clinical randomized trial (CRT), the blood level of LDH was selected as an inclusion criterion. However, LDH decreased during the pandemic; hence, the impact of this decrease on the prognostic value of LDH for mortality was evaluated. Methods: Data on LDH levels in 843 patients were obtained and analyzed. Relative risk, standard error and receiver operating characteristic curves were calculated for two cutoff values. Results: Relative risk lost validity and the area under the curve narrowed by trimester during the pandemic. Conclusion: The progressive decrease in LDH impacted the capacity to predict mortality in COVID-19. More studies are needed to validate this finding and its implications.
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COVID-19 , L-Lactato Desidrogenase , Humanos , COVID-19/enzimologia , COVID-19/epidemiologia , L-Lactato Desidrogenase/metabolismo , Pandemias , Prognóstico , Estudos Retrospectivos , Curva ROCRESUMO
Alcohol consumption is an issue of worldwide relevance and a problem of national scale in Mexico. The consumption pattern of large amounts of alcohol on the weekends is rapidly increasing in young adults between 18 and 29 years. Despite various studies that have focused on the noxious effect of alcohol in immunity, the changes in the immunoprofiles of peripheral blood cells have not been completely described. Natural killer cells (NKCs) are lymphoid-origin cells of the immune system that are responsible for defense against tumors, among other functions. In homeostatic conditions, they are found to be in a state of "dynamic balance" between activation and inhibition stimuli, which, if broken, may lead to immunosuppression or activation of cytotoxic mechanisms. In this study, we evaluated the immunoprofile of peripheral NKCs of 54 young adults, 29 of whom were binge drinkers and 25 of whom were low risk (LR), as classified by validated tools. Drinking habits were assessed. Blood samples were collected to perform hematic biometry and liver enzyme tests. Peripheral NKCs were identified by FACS, and stained for CCR2, CCR4, CCR5, CXCR4, CD69, CD127, CD137, TLR4, and Granzyme B. The data were analyzed using the t test and Mann-Whitney's U test for contrasts, and the effect size was obtained in order to evaluate the impact of each immunoprofile. The binge group showed increased expression of CCR5 and PD-1 in NKCs, respective to the LR group, and decreased expression of TLR4, along with fewer CCR4+ cells. Moreover, the increase found in CCR5 and PD-1 expression was correlated with the number of drinks in the last drinking session. Our findings show that young binge drinkers have different immunoprofiles that could suggest an early status of immunosuppression and trafficking of NKCs to the liver, which could be related to the onset of early liver damage, early in a subject's lifespan.